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The application of alternative methods for skin irritation evaluation on pesticides (농약에 대한 피부자극성 대체시험법 적용연구)

  • Jeong, Mi-Hye;Kim, Mi-Kyoung;Park, Soo-Jin;You, Are-Sun;Hong, Soon-Sung;Park, Kyung-Hun;Park, Jae-Eup
    • The Korean Journal of Pesticide Science
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    • v.16 no.3
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    • pp.261-266
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    • 2012
  • It is common to use many experiment animals to evaluate the toxicity of chemicals including pesticides. For protecting animal, the concepts of 3R (Reduction, Replacement, Refinement) were introduced and in vitro alternatives methods actively have been developed all over the world. Many experimental animals for toxicological tests have been used, so that it is important to establish the alternative methods. In this study, the alternative method using reconstituted human skin model (Keraskin$^{TM}$) was conducted for classification of skin irritation on pesticides. Sixteen formulations selected on the basis of the degree of irritation were treated by Keraskin$^{TM}$ test. The percent of cell viability was measured into the culture medium collected after treatment of the pesticides for 24-72 hrs. The skin irritations of formulations were evaluated by the cell viability. In this study, The 4 formulations with mild irritation in rabbits were evaluated as nonirritant, the 6 formulations with moderate and severe irritation were evaluated as irritant in human skin model test. We suggest that the alternative test using Keraskin$^{TM}$ model could be used as toxicity evaluation for primary irritation index (P.I.I.) score of greater than or equal to 2.1 of pesticides. The further studies should be required to apply for hazardous assessment of pesticides on alternative skin irritation methods because of the interindividual variability of the sensitivity of skin irritation on pesticides.

Inhibitory Effects of Asparagus cochinchinensis in LPS-Stimulated BV-2 Microglial Cells through Regulation of Neuroinflammatory Mediators, the MAP Kinase Pathway, and the Cell Cycle (Lipopolysaccharide로 자극된 BV-2 미세교세포에서 신경염증 매개체, MAP kinase경로, 세포주기의 조절에 의한 천문동(Asparagus cochinchinensis)의 저해효과)

  • Lee, Hyun Ah;Kim, Ji Eun;Choi, Jun Young;Sung, Ji Eun;Youn, Woo Bin;Son, Hong Joo;Lee, Hee Seob;Kang, Hyun-Gu;Hwang, Dae Youn
    • Journal of Life Science
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    • v.30 no.4
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    • pp.331-342
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    • 2020
  • The suppression of neuroinflammatory responses in microglial cells can be considered a key target for improving the progression of neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). Asparagus cochinchinensis has traditionally been used as a medicine to treat fever, cough, kidney disease, breast cancer, inflammatory diseases, and brain diseases. In this study, we investigated the neuroprotective mechanism of an aqueous extract from A. cochinchinensis root (AEAC), particularly its anti-inflammatory effects on lipopolysaccharide (LPS)-activated BV-2 microglial cells. BV-2 cells were treated with four different concentrations of AEAC. No significant toxicity was detected in BV-2 cells treated with AEAC. Nitric oxide (NO), cyclooxygenase-2 (COX-2) mRNA, and inducible nitric oxide synthase (iNOS) mRNA levels were 21% lower in the AEAC+LPS group than in the Vehicle+LPS group. Lower proinflammatory (TNF-α and IL-1β) and anti-inflammatory cytokine (IL-6 and IL-10) levels were also detected in the AEAC+LPS group than in the Vehicle+LPS group, albeit at varying rates. Moreover, the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group compared to the Vehicle+LPS group, enhancement of the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group, while cell cycle arrest at the G2/M phase caused by LPS treatment was less severe in the AEAC+LPS group. The increase in reactive oxygen species (ROS) generation induced by LPS treatment was also lower in the AEAC-pretreated group than in the Vehicle+LPS group. This is the first study to show that AEAC exerts anti-neuroinflammatory activity against LPS stimulation by regulating the MAPK signaling pathway, the cell cycle, and ROS production.

Coronary Artery Bypass Grafting with Cardiopulmonary Bypass Versus Without Cardiopulmonary Bypass (체외순환 및 비체외순환하에서의 관상동맥우회로술)

  • Park, Chan-Beom;Kwon, Jong-Bum;Park, Kuhn;Won, Yong-Soon
    • Journal of Chest Surgery
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    • v.34 no.8
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    • pp.591-596
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    • 2001
  • Coronary artery bypass graft with cardiopulmonary bypass is a conventional method of operative revascularization of coronary artery disease. Because of many troubles of cardiopulmonary bypass such as systemic inflammatory reaction, mechanical trauma of blood components and coagulopathy, coronary artery bypass graft without cardiopulmonary bypass has been popularized. Material and Method: From March 1999 to September 2000, 35 patients under went CABG at our institution. Among them, 14 patients received CABG without the use of CPB and 21 patients under went CABG with the use of CPB. Mean operative time, mean postoperative tracheal intubation time, mean ICU stay, mean hospital stay, the amount of transfusion, postoperative use of inotropic agents, and postoperative changes of cardiac enzymes were compared in both groups. Result: There were differences between the CABG without CPB group and CABG with CPB group with regard to mean tracheal intubation time, the amount of transfusion and the elevation of postoperative cardiac enzymes(p<0.05). Conclusion: While CABG without CPB provided satisfactory results, more long term follow-up is required.

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Studies on Molecular Plasticity of Bergmann Glia following Purkinje Cell Degeneration (조롱박신경세포의 변성에 따른 버그만아교세포의 면역조직학적 연구)

  • Yoon, Chul-Jong;Cho, Sa-Sun;Lee, Ha-Kyu;Park, Min-Chul
    • Applied Microscopy
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    • v.35 no.3
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    • pp.165-176
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    • 2005
  • Studies on molecular plasticity of Bermann glia (BG) after harmaline-induced Purkinje cell (PC) degeneration in the rat cerebellum. The intimate structural relationship between BG and PC, evidenced by the sheathing of the PC dendrites by veil-like process from the BG has been suggestive of the close functional relationship between these two cell types. However, little is known about metabolic couplings between these cells. This study designed to investigate molecular plasticity of BG in the rat cerebellum in which PCs were chemically ablated by harmaline treatment. Immunohistochemical examination reveals that harmaline induced PC degeneration causes a marked glial reaction in the cerebellum with activated BG and microglia aligned in parasagittal stripes within the vermis. In these strips, activated BG were associated with upregulaion of metallotheionein, while GLAST and was down regulated, as compared with nearby intact area where both BG are in contact with PCs. The data from this study demonstrate that BG can change their phenotypic expression when BG loose their contact with PCs. It is conceivable that activated BG may upregulate structural proteins, metallothionein expression to use for their proliferation and hypertrophy; metallothionein expression to cope with oxidative stress induced by PC degeneration and microglial activation. On the contrary, BG may down regulated expression of GLAST because sustained loss of contact with PCs would eliminate the necessity for the cellular machinery involved glutamate metabolism. In conclusion, BG might respond man to death of PCs by undergoing a change in metabolic state. It seems possible that signaling molecules released from PCs regulates the phenotype expression of BG. Also ultrastructures in the organelles of normal PC and BG are distinguished by mitochondrial appearance, and distributed vesicles at the synaptic area in the cytoplasm.

Anti-corrosion properties for cross section of Mg films on galvalume steel coated by PVD process (PVD법에 의해 Mg 코팅된 갈바륨 도금강판의 단면부 내식특성)

  • Park, Jae-Hyeok;Kim, Sun-Ho;Park, Gi-Dong;Jeong, Jae-In;Yang, Ji-Hun;Lee, Gyeong-Hwang;Lee, Myeong-Hun
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2018.06a
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    • pp.65-65
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    • 2018
  • 갈바륨 도금강판은 알루미늄의 우수한 차폐 특성과 내열성, 열 반사성을 가지며 아연의 희생방식 특성을 겸비하여 동일 부착량의 용융 아연도금 및 알루미늄 도금강판에 비해 우수한 내식성을 나타낸다고 알려져 있다. 또한 이것은 표면이 미려하고 경제성이 높아 건자재 용도로 현재까지도 세계적으로 널리 이용되고 있다. 여기서 지칭하는 바륨 도금강판(galvalume steel)은 아연과 알루미늄 도금강판의 장점을 접목하기 위해 55 Al-43.4 Zn-1.6 Si (wt.%)로 구성되어 개발된 3원계 성분의 합금도금강판이다. 한편, 최근에는 강재의 내식성을 향상시키기 위한 다양한 연구 결과에 의해 Zn-Al-Mg의 3원계 합금도금강판도 개발되어 사용되고 있다. 이것은 기존의 아연도금 강판 보다 10배 정도의 우수한 내식성을 나타내는 것으로 보고되고 있다. 특히, 이것은 도금된 평판부의 내식성은 물론 절단된 도금 단면부의 내식성도 매우 우수하다고 알려져 있다. 그러나 상기한 갈바륨 도금강판의 경우에는 도금된 표면부에 비해 단면부의 내식성이 상대적으로 취약한 것으로 알려져 있다. 따라서 본 연구에서는 갈바륨 도금강판의 내식성을 종합적으로 향상시키기 위하여 이 갈바륨 도금강판 상에 PVD 스퍼터링법에 의해 Mg 코팅막의 제작을 시도하였다. 여기서 Mg 코팅막은 후처리 된 갈바륨 도금강판 상에 Ar 공정압력 2 및 20 mTorr 조건 중 1.5 및 $3{\mu}m$ 두께로 제작하였다. 또한 제작한 코팅막에 대해서는 모폴로지 관찰(SEM) 및 결정구조 분석(XRD)을 하였고, 분극측정, 염수분무 시험(SST) 및 복합부식 시험(CCT)에 의해 표면 및 단면부의 내식성평가를 수행하였다. 또한 여기서는 염수분무 및 복합부식 시험 후의 시험편도 채취 하여 표면 및 단면부위에 대한 원소조성 분석(EPMA)과 결정구조 분석(XRD)을 실시하였다. 이상의 실험 결과에 의하면, 본 실험에서 제작한 Mg 코팅막은 그 두께가 두꺼울수록 표면 Mg 결정립의 크기가 증가하였고, 그 부식속도가 증가하는 경향을 나타내었다. 또한 여기서는 공정압력이 높은 조건에서 제작한 막일수록 Mg(002)면 피크 강도가 감소하고 Mg(101)면 피크의 배향성이 증가하였다. 그때 그 막의 내식성은 향상되는 경향을 나타내었다. 그리고 종합적으로 염수분무 및 복합부식 시험 결과에 의하면 Mg이 코팅된 갈바륨 도금강판은 기존 갈바륨 도금강판 보다 내식성이 현저히 향상되었다. 특히, 단면부 내식성의 경우에는 기존 대비 5배 이상 향상되는 경향을 나타내었다. 여기서 단면부 내식특성 분석을 위한 EPMA 원소조성 분석 결과에 의거하면, 부식 초기에는 마그네슘의 부식생성물에 의해 단면부가 치밀하게 보호되고 있음을 확인할 수 있었다. 그 이후에는 부식이 지속적으로 진행됨에 따라 갈바륨 도금층에서 용출된 알루미늄 및 아연 성분이 마그네슘과 함께 치밀한 부식생성물을 형성하여 단면부를 차폐함에 따라 단면부의 내식성이 크게 향상된 것으로 생각된다. 이러한 부식생성물의 결정구조 분석 결과에 따르면, 염수분무와 복합부식 시험에서는 공통적으로 MgO, $Mg(OH)_2$ 이외에도 Simonkolleite상 등이 형성되었다. 또한 건-습 반복 부식시험인 복합부식시험 후에는 $Mg_5(CO_3)_4(OH)_24H_2O$(Hydromagnesite)상 등이 형성됨을 확인할 수 있었다. 즉, 본 실험에서 후처리된 갈바륨 도금강판 상에 제작한 마그네슘 코팅막의 경우에는 상기와 같은 다양한 부식반응에 의해 표면 및 단면부에 형성된 Mg계 부식생성물과 $Zn_5(OH)_8Cl_2H_2O$(Simonkolleite)상에 의해서 표면은 물론 단면부 내식성이 크게 향상된 것으로 사료된다.

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The Effect of LhGH on Hair Regeneration in C57BL/6CrN Mouse (LhGH가 마우스(C57BL/6CrN)의 모발 재성장에 미치는 영향)

  • Kim, Yong-Ju;Kim, Tae-Keun;Min, Byoung-Hoon;Kim, Soo-Jin
    • Applied Microscopy
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    • v.41 no.1
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    • pp.47-53
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    • 2011
  • Hair is an appendage of skin which protects the body from outer physical and chemical stimuli. Hair is generated from the hair follicle lying on a sunken basal layer of epidermis. Hair cycling, which regenerates hair follicles throughout the life time of the organism. Numerous kinds of factors which exist at the hair follicle have been reported to regulate hair cycling, Human growth hormone secreted from pituitary gland, initially demonstrated to accelerate organ's growth, has been reported to play a role in the biology of organ size determination. We investigated the effect of 6-histidines residues tagged at amino-terminus of human growth hormone using light and electronmicroscopic methods. Human growth hormone encapsulated in nano-liposome (LhGH) was used to find how LhGH affects hair follicle cycling of mouse (C57BL6/CrN). Distilled water as a negative control, 3% Minoxidil as a positive control, and LhGH were applied to mouse for weeks. LhGH increased the number of exposed hairs per given areas ($1mm^2$). This result was also confirmed using a different breed of mice which show natural hair loss in an old age (about 17 months after birth). When LhGH was applied for 3 weeks after natural hair loss, natural hair loss on these mice was prevented, However, the control group mice on which LhGH was not applied showed further hair loss. This result indicates that LhGH may stimulate hair cycling of mouse. In clusion, it is cleat that the LhGH increased the number of hair on mice and help the depilated skin to grow new hair follicles again.

Inhibitory Effects of Schisandrae Fructus Ethanol Extract on the Production of Matrix Metalloproteinases in in vitro and in vivo Osteoarthritis Models (In vitro 및 in vivo 퇴행성관절염 모델에서 오미자 에탄올 추출물에 의한 matrix metalloproteinases의 생성 억제)

  • Jeong, Jin-Woo;Lee, Hye Hyeon;Kim, Hong Jae;Lee, Ki Won;Kim, Ki Young;Kim, Sung Goo;Hong, Su-Hyun;Kim, Bum Hoi;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1207-1214
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    • 2017
  • Schisandrae Fructus (SF), the fruit of Schisandra chinensis (Turcz.) Baill., is widely used in traditional medicine for the treatment of a number of chronic diseases. SF extracts have been recently reported to attenuate the inflammatory responses in SW1353 human chondrocyte cells in in vitro and monosodium iodoacetate (MIA)-induced cartilage degradation in in vivo osteoarthritis (OA) models. However, their protective and therapeutic potentials against OA in primary culture chondrocytes and animal models remain unclear. Therefore, we investigated the effects of the ethanol extract of SF on the activity of matrix metalloproteinases (MMPs), biomarkers for diagnosis of OA, on interleukin $(IL)-1{\beta}-induced$ primary cultured rat cartilage chondrocytes and MIA-induced osteoarthritis in a rat model. Our data indicated that SF treatment significantly reduced the mRNA expression and enzyme activity of MMP-1, -3 and -13 in $IL-1{\beta}-induced$ primary cultured rat cartilage chondrocytes. The chondro-protective effects of SF were then analyzed in a rat OA model using a single intra-articular injection of MIA in the right knee joint. According to our results, the elevated levels of MMP-1 and -3 were markedly ameliorated by SF administration. Collectively, these findings indicate that SF could be a candidate for the treatment of OA.

Comparative Chemical Compositions of Four Kinds of Tochukaso (동충하초 품종별 영양성분 비교)

  • Oh, Se-Wook;Kim, Sun-Hee;Song, Hyo-Nam;Han, Dae-Seok
    • Korean Journal of Food Science and Technology
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    • v.35 no.1
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    • pp.15-22
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    • 2003
  • Nutritional compositions of three Tochukaso species (Paecilomyces tenuipes hosted by Larva and pupa, Cordyceps militaris, C. sinensis) were compared. Fruiting body and host fractions were separately analyzed. Fruiting body fraction of P. tenuipes (36.6%) hosted by larva was higher than that hosted by pupa (10.2%), an indication that the quality of the former is superior to the latter. Carbohydrate content of C. sinensis (39.6%) was $2.5{\sim}7$ times higher than those of others, probably due to the presence of polysaccharides. Protein and crude lipid contents of C. sinensis and C. militaris were 25.8 and 10.3%, and 75.1 and 3.9%, respectively. C. sinensis showed the lowest Ca content and $30{\sim}75$ times higher Fe content among the samples tested. Vitamin A content of C. militaris was 308.9 IU/100g, two fold higher than those of the other species. Saturated fatty acid content was the highest in P. tenuipes (pupa, 27.7%), whereas unsaturated fatty acid was the highest in P. tenuipes (larva, 83.3%). Aspartic acid, glutamic acid, and glycine were abundant in all species. Cordycepin content of C. militaris was $20{\sim}50$ times higher than those of the other species.

A Study on Anti-Bone Resorption & Osteoporosis by Taeyoungion-Jahage Extracts

  • Bae, Hyo-Sang;Cho, Hyung-Lae;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Shin, Jung-Sik
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.61-75
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    • 2002
  • Recombinant human $interleukin-1{\beta}$ $(rhIL-1{\beta})$ regulates several activities of the osteoblast cells derived from mouse calvarial bone explants in vitro. $rhIL-1{\beta}$ stimulated cellular proliferation and the synthesis of prostaglandin $E_2(PGE_2)$ and plasminogen activator activity in the cultured cells in a dose-dependent manner. However, the induction of osteocalcin synthesis and alkaine phosphatase activity in response to vitamine D, two characteristics of the osteoblast phenotype, were antagonized by $rhIL-1{\beta}$ over a similar dose range. This study supports the role of $IL-1{\beta}$ in the pathological modulation of bone cell metabolism, with regard to implication in the pathogenesis of osteoporosis by $IL-1{\beta}$. When the mouse calvarial bone cells were used, the bone resorption induced by $IL-1{\beta}$ was strongly inhibited by calcitonin treatment, indicating osteoclast-mediated bone resorption. On the other hand, the medicinal extracts of Taeyoungjon-Jahage (T.Y.J-J.H.G extracts) was tested for whether they could inhibit $IL-1{\beta}-induced$ $PGE_2$ production. Cell viability was not significantly affected by treatment with the indicated concentration of the extracts. The T.Y.J.-J.H.G. extracts were shown to have the inhibitory effects against the synthesis of $PGE_2$. We also examined the effect of the pretreatment with a various concentrations of the T.Y.J.-J.H.G. extracts then treated the $PGE_2-induction$ agents. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h, which by itself had little effect on cell survival, did not enhance the synthesis of $PGE_2$. Furthermore, the T.Y.J-J.H.G. extracts were shown to have the protective effects against plasminogen dependent fibrinolysis induced by the bone resorption agents of $IL-1{\beta}$. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h did not enhance the plasminogen dependent fibrinolysis. Finally, calcitonin showed the inhibitory activity the $IL-1{\beta}-stimulated$ bone resorption in the mouse calvarial bone cells having both of the osteoblast and osteoclast cells. Seemingly, pretreatment of the T.Y.J.-J.H.G. extracts for 1 h reduced the bone resorption. These results clearly indicated that calcitonin and T.Y.J.-J.H.G. extracts play key roles in inhibition of the osteoclast-mediated bone resorption.

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Noninvasive Prenatal Diagnosis using Cell-Free Fetal DNA in Maternal Plasma: Clinical Applications

  • Yang, Young-Ho;Han, Sung-Hee;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.1-16
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    • 2011
  • Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.